cholestyramine resin  (Macklin Inc)

Journal: Microbiome

Article Title: Maternal Western diet mediates susceptibility of offspring to Crohn’s-like colitis by deoxycholate generation

doi: 10.1186/s40168-023-01546-6

Figure Lengend Snippet: MWD promotes colitis development in their progeny by producing DCA. a As shown in Fig. a, WT mice ( n = 5) were colonized with gut microbiota from the W-N and N-N groups. Then, the concentration of DCA in the mice’s feces was measured using an ELISA assay kit. b – g Cholestyramine resin (resin) was given to mice in the W-N and N-N groups ( n = 6 per group) for 5 days to eliminate intestinal bile acids. Following that, mice were given TNBS to induce colitis. c Body weight changes in mice were evaluated daily after TNBS treatment. d Representative images of TNBS-treated colon in N-N+resin and W-N+resin groups. e The mice were sacrificed on day 4, and the colon length was recorded. f , g Histopathological analysis of colon sections. f Histological scores of colitis were assessed. g Representative images of the H&E-stained colon sections of relevant groups (scale bars 100 μm). a , c , e , and f Data represent means ± SEM; NS, not significant; ** P < 0.01; by unpaired Student’s t test. The data shown are representative of three independent experiments

Article Snippet: Cholestyramine resin (11,041–12-6) was obtained from MedChem Express.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Staining

Journal: Journal of Pharmaceutical Analysis

Article Title: Traditional Chinese medicine Pien-Tze-Huang ameliorates LPS-induced sepsis through bile acid-mediated activation of TGR5-STAT3-A20 signalling

doi: 10.1016/j.jpha.2023.12.005

Figure Lengend Snippet: Bile acid (BA), which is an important functional component in Pien Tze Huang (PZH), regulates activation of the G protein-coupled bile acid receptor 1 (TGR5)-signal transducer and activator of transcription 3 (STAT3)-A20 signalling pathway. (A) Total BA levels were detected in the PZH solution and PZH plus cholestyramine resin. (B) Liquid chromatography-mass spectrometry (LC-MS) analysis of PZH supernatant treated with or without cholestyramine resin. (C) Peritoneal macrophages were pretreated with PZH (1.25 mg/mL) or cholestyramine resin-treated PZH (1.25 mg/mL) for 2 h, and then treated with lipopolysaccharide (LPS) (100 ng/mL) for 40 min. PZH treated with cholestyramine resin did not increase the protein expression of TGR5, phospho-STAT3 (p-STAT3) and A20 or decrease the protein expression of phospho-p65 (p-p65), phospho-extracellular signal-regulated kinases (p-ERK), and phosphor-C-Jun N-terminal kinases (p-JNK) induced by LPS. (D) Peritoneal macrophages were pretreated with PZH (1.25 mg/mL) or cholestyramine resin-treated PZH (1.25 mg/mL) for 2 h and then treated with LPS (100 ng/mL) for 6 h. PZH treated with cholestyramine resin did not decrease the mRNA levels of interleukin (IL)-6, tumour necrosis factor-α (TNF-α) and IL-1β induced by LPS. (E) The composition of BAs in the PZH solution was detected by LC-MS. (F) Peritoneal macrophages were pretreated with PZH (1.25 mg/mL), cholestyramine resin-treated PZH (1.25 mg/mL), and cholestyramine resin-treated PZH (1.25 mg/mL) + BAs for 2 h, respectively, and then treated with LPS (100 ng/mL) for 6 h. Replenishing BAs including cholic acid (CA) (61.53 μM), deoxycholic acid (DCA) (27.25 μM), chenodexycholic acid (9.75 μM), taurocholic acid (TCA) (20.52 μM), glycocholic acid (GCA) (7.466 μM) and lithocholic acid (LCA) (0.413 μM) in PZH after treatment with cholestyramine resin decreased the mRNA levels of TNF-α and IL-1β induced by LPS. (G) Peritoneal macrophages were pretreated with PZH (1.25 mg/mL), cholestyramine resin-treated PZH (1.25 mg/mL), and cholestyramine resin-treated PZH (1.25 mg/mL) + BAs for 2 h and then treated with LPS (100 ng/mL) for 40 min . Replenishing BAs including CA (61.53 μM), DCA (27.25 μM), chenodexycholic acid (9.75 μM), TCA (20.52 μM), GCA (7.466 μM) and LCA (0.413 μM) in PZH after treatment with cholestyramine resin decreased the expression of p-p65 and p-ERK induced by LPS. (H) Schematic model of the mechanism by which PZH attenuates the LPS-induced inflammatory response by activating TGR5-STAT3-A20 signalling pathway. PZH stimulates the phosphorylation of STAT3, which acts as a transcriptional activator for A20 to increase the expression of A20. A20 inhibits the activation of the nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) signalling pathways to decrease the production of proinflammatory cytokines to attenuate the sepsis progression. The results are representative of one of three experiments. The data are shown as the mean ± standard deviation (SD). ∗∗ P < 0.01, ∗∗∗ P < 0.001, ns: no significance, based on one-way analysis of variance (ANOVA) and two-tailed Student's t -test. CHOL: cholestyramine resin; TRAF6: tumor necrosis factor receptor-associated factor 6; IKK: nuclear factor- kappa B kinase; TAK1: transforming growth factor beta-activated kinase 1; IκBα: nuclear factor-kappa B inhibitor alpha.

Article Snippet: Cholestyramine resin was purchased from Shanghai Yuanye Bio-Technology Co., Ltd (Shanghai, China).

Techniques: Functional Assay, Activation Assay, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Expressing, Phospho-proteomics, Standard Deviation, Two Tailed Test